The effectiveness of insulin therapy in the functional and morphological recovery of leydig cells in rats submitted to Diabetes mellitus

The effects of systemic insulin administration at different concentrations on the testicular tissue of diabetic adult rats, induced by streptozotocin, are evaluated by the morphological analysis of spermatogenic process. Twenty-four adult male rats were divided into 1) Control Group: they received citrate buffer, by intraperitoneal injection; 2) Diabetic Group: induced by intraperitoneal injection of streptozotocin (60 mg. kg-1 of body weight); 3) Insulin 50%: induced diabetes treated with half of standard dosage of insulin; 4) Insulin 100%: induced diabetes treated with standard dose of insulin. After eight weeks, animals were weighted and anesthetized; testicles were removed and processed in resin. Body and testicular weight of diabetic rats decreased when compared to that of control. Parameters increased with insulin therapy. Testosterone levels were low in diabetic animals but rates recovered after insulin therapy. Nuclear diameter and volume of Leydig cells decreased in diabetic rats although they significantly increased after insulin therapy. Results showed that the administration of insulin in diabetic rats promoted a protective effect of testicular parenchyma, enhancing efficient recovery on testosterone levels and increase in daily sperm production.

Testicular histological evaluation and serum testosterone concentrations of bulls after chemical castration with calcium chloride / Avaliação histológica e das concentrações séricas de testosterona de touros após a castração química com cloreto de cálcio

Orchiectomized bulls have advantages in the meat quality and ease of handling. Chemical castration is an option for surgical castration and the sclerosing agents can be administered into the testicular or epididymis parenchyma. These agents have a lower incidence of complications than surgery, especially when associated with dimethylsulfoxide (DMSO), which has anti-inflammatory action and increases the absorption of other drugs. Thus, this study aimed to evaluate the effect of a single intratesticular injection of calcium chloride solution associated with DMSO for the chemical sterilization of bulls. Twenty-four young adult bulls were utilized, distributed into 3 groups (G20, G30 and G40, n = 8/group), according to the calcium chloride concentration (20, 30 and 40%), in 10mL volume. Serum concentrations of testosterone, body weight, testicular volume and ecotexture, clinical signs and behavior and were evaluated for 45 days. Thus, the animals were orchiectomized and testicles were assessed histologically. There were no changes in body weight, decreased serum testosterone concentrations (except G30), signs of scrotal sensitivity or changes in behavior over the period. However, there was significant increase in testicular volume, especially on the 2nd and 3rd day after treatment, with values returning to the value initials at 15 days. Testicular adherence and firm consistency were observed during orchiectomy. Ultrasound examination revealed a loss of integrity of the median raphe, with cavity formation and an alteration of the testicular echotexture. In the histological evaluation, coagulation necrosis of seminiferous tubules and interstitial cells was observed, mainly in the medial portion in all groups. Some animals presented total absence of tubular formations in all the studied groups, being the effects of greater intensity in the G40. Additionally, pronounced edema was noted in all groups, especially in G40. Inflammatory infiltrate, fibroplasia and neovascularization were found to be predominantly discrete. Based on the conditions used in this study, we conclude that calcium chloride associated with DMSO can be used as a method of chemical sterilization in bovines.(AU)

Bovinos orquiectomizados apresentam vantagens na qualidade da carne e facilidade no manejo. A quimioesterilização é uma opção à castração cirúrgica e os agentes esclerosantes podem ser administrados no parênquima testicular ou epidídimo. Estes produtos possuem menor incidência de complicações, comparados a cirurgia, especialmente quando associados ao dimetilsulfóxido (DMSO), que apresenta ação anti-inflamatória e aumenta a absorção de outros fármacos. Assim, este estudo teve como objetivo avaliar o efeito de uma única injeção intratesticular de solução de cloreto de cálcio associado com 0,5% de DMSO para a esterilização química de bovinos. Vinte e quatro touros adultos jovens foram utilizados, distribuídos em 3 grupos (G20, G30 e G40, n = 8/grupo) de acordo com a concentração de cloreto de cálcio (20, 30 e 40%), em um volume de 10mL. Foram avaliadas as concentrações séricas de testosterona, peso corporal, volume e ecotextura testicular, sinais clínicos e comportamento por 45 dias. A seguir, os animais foram submetidos à orquiectomia e os testículos avaliados histologicamente. Não foram observadas alterações do peso corporal, diminuição das concentrações de testosterona sérica (exceto no G30), sinais de sensibilidade escrotal ou alterações no comportamento no período avaliado. Porém, houve aumento significativo do volume testicular, especialmente nos 2º e 3º dia após o tratamento, com valores retornando aos iniciais aos 15 dias. Aderência e consistência firme dos testículos foram achados observados durante a orquiectomia. O exame ultrassonográfico revelou perda de integridade da rafe mediana, com formação de cavidades e alteração da ecotextura testicular. Na avaliação histológica, verificou-se necrose de coagulação de túbulos seminíferos e células intersticiais acentuada, principalmente, na porção medial em todos os grupos, sendo que em alguns animais havia ausência total das formações tubulares em todos os grupos estudados, sendo os efeitos de maior intensidade no G40. Além disso, edema foi acentuado em todos os grupos, principalmente em G40. Infiltrado inflamatório, fibroplasia e neovascularização foram achados predominantemente discretos. Com base nas condições utilizadas neste estudo, conclui-se que o cloreto de cálcio associado com o DMSO pode ser utilizado como um método de esterilização química em bovinos.(AU)

Testicular Histomorphometric Evaluation of Zebu Bull Breeds

The objective of this study was to evaluate the quantitative histology and testicular biometrics in zebu bulls of different breeds. Testicular fragments of Nelore (n=10), Polled Nelore (n=6), Gir (n=5), Guzerat (n=5) and Tabapuã bulls (n=5) were used. The fragments were perfusion-fixed in Karnovsky solution, embedded in glycol methacrylate and stained with toluidine blue-1% sodium borate. The Nelore animals had a higher tubular volumetric proportion (85.2%) and greater height of the seminiferous epithelium (73.2 µm) than the Gir, Guzerat and Tabapuã breeds. The Nelore animals also had a higher volumetric proportion of Leydig cells (5.2%) than the Guzerat and Tabapuã breeds. There was no significant difference for any of these parameters between the Nelore and Polled Nelore breeds. The gonadosomatic index, seminiferous tubule diameter, cross-sectional area of the seminiferous tubule and tubule length (total length and length per gram of testicular parenchyma) did not vary among the breeds studied. The morphometric parameters evaluated suggested that the genetic selection applied to the Nelore and Polled Nelore breeds improved the efficiency of spermatogenesis in these breeders.

Differential expression of speckled POZ protein, SPOP: Putative regulation by miR-145.

The speckle POZ protein, SPOP, is an adaptor of the Cul3-based ubiquitination process, and has been implicated in the carcinogenesis process. Despite recent elucidation of biological functions, regulation of SPOP gene expression has not been reported. In this study, the mRNA levels of the mouse SPOP (mSPOP) gene were first shown to vary noticeably in different tissues. However, the SPOP protein was detected in high abundance only in Purkinje cells of the cerebellum and seminiferous tubule of the testis, echoing previous reports of involvement of ubiquitination in neuron cells and in spermatogenesis. In other mouse tissues and human cancer cell lines analysed, only low SPOP protein levels were detected. The 3′-untranslated regions of both the mSPOP and human SPOP transcripts harbor a conserved putative miR-145 binding site (BS). In some tissues and cell lines, miR-145 and SPOP protein levels were in an inverse relationship suggesting miR-145 regulation. Luciferase assays of deletion and point mutation constructs of the miR-145 BS, and miR-145 induction by serum starvation that resulted in reduced endogenous SPOP levels provided further evidence that miR-145 is likely involved in post-transcriptional regulation of SPOP expression in selected tissues, and possibly with the participation of other miRNA species.

Differential expression of speckled POZ protein, SPOP: Putative regulation by miR-145.

The speckle POZ protein, SPOP, is an adaptor of the Cul3-based ubiquitination process, and has been implicated in the carcinogenesis process. Despite recent elucidation of biological functions, regulation of SPOP gene expression has not been reported. In this study, the mRNA levels of the mouse SPOP (mSPOP) gene were first shown to vary noticeably in different tissues. However, the SPOP protein was detected in high abundance only in Purkinje cells of the cerebellum and seminiferous tubule of the testis, echoing previous reports of involvement of ubiquitination in neuron cells and in spermatogenesis. In other mouse tissues and human cancer cell lines analysed, only low SPOP protein levels were detected. The 3′-untranslated regions of both the mSPOP and human SPOP transcripts harbor a conserved putative miR-145 binding site (BS). In some tissues and cell lines, miR-145 and SPOP protein levels were in an inverse relationship suggesting miR-145 regulation. Luciferase assays of deletion and point mutation constructs of the miR-145 BS, and miR-145 induction by serum starvation that resulted in reduced endogenous SPOP levels provided further evidence that miR-145 is likely involved in post-transcriptional regulation of SPOP expression in selected tissues, and possibly with the participation of other miRNA species.

Unilateral cryptorchidism induces morphological changes of testes and hyperplasia of Sertoli cells in a dog / 한국실험동물학회지

Cryptorchidism is one of the most common genital defects in dogs. This study investigated the effects of abdominal cryptorchidism on morphology, cell proliferation, and Sertoli cell condition in a dog with spontaneous unilateral cryptorchidism. Elective orchidectomy was performed on the abdominal right testis and the scrotal left testis. Significant reductions in numbers of spermatogonia, spermatocytes, and spermatids were observed in hematoxylin and eosin stained sections of the cryptorchid testis. The size of the epididymal duct was smaller than that of the control testis. Based on Ki67 immunohistochemistry, the proliferative activity of spermatogonia and spermatocytes was significantly decreased in the cryptorchid testis. However, proliferative activity was increased in the epididymal duct. Based on GATA-4 immunohistochemistry, Sertoli cells were relatively resistant to cryptorchidism, and the proliferative activity of Sertoli cells was markedly increased in the cryptorchid testis than in the control testis. These results suggest that spontaneous unilateral cryptorchidism causes morphological defects in spermatogonia and spermatocytes in the testis and changes the size of the efferent ductule of the epididymis. In addition, spontaneous unilateral cryptorchidism increases proliferative activity of Sertoli cells, which may be a predisposing factor for Sertoli cell cancer in cryptorchid testes.

Análise morfológica e funcional do processo espermatogênico em cobaios (Cavia porcellus) da pré-puberdade até a pós-puberdade / Morphological and functional analysis of spermatogenesis in guinea pigs (Cavia porcellus) from pre-puberty to post-puberty

Este estudo descreveu as análises morfológica e funcional do processo espermatogênico em cobaios (Cavia porcellus) de cinco (S5); seis (S6); nove (S9) e onze (S11) semanas de idade (N=5/grupo). Os aspectos analisados incluíram a contagem das populações celulares presentes no estádio 1 do ciclo do epitélio seminífero (CES), eficiência das mitoses espermatogoniais (RMi), produção meiótica (RMe), rendimento geral da espermatogênese (RGE), índice de células de Sertoli (ICS) e capacidade de suporte das células de Sertoli (CSCS). Os resultados mostraram que número médio de espermatogônias A, espermatócitos primários em pré-leptóteno/leptóteno, espermatócitos primários em paquíteno, células espermatogênicas totais e células de Sertoli mostraram variações numéricas em função da idade, entretanto, não detectadas estatisticamente, enquanto espermátides arredondadas aumentaram significativamente na puberdade e depois se estabilizaram. A produção espermatogênica de cobaios de 5 a 11 semanas não atingiu o ponto de estabilização e o RMi, RMe, RGE, ICS e CSCS mostraram variação numérica significativa em função da idade. Os resultados demonstraram que Cavia porcellus na pós-puberdade 2 são um modelo experimental vantajoso para estudos de processos de reconhecimento homólogos, alinhamento, e sinapses durante a prófase meiótica; o rendimento intrínseco da espermatogênese em cobaios é semelhante ao relatado para ratos Wistar, pacas e cutias (Dasyprocta sp.) e menor do que em preás, enquanto que a eficiência funcional das células de Sertoli é superior a de cutias e ratos Wistar e inferior à de pacas, rato espinhoso e catetos. Concluiu-se que em cobaios a espermatogênese está completamente estabelecida na semana 6 de idade, indicando a fase púbere do desenvolvimento sexual, e até a semana 11 eles não atingiram a produção espermática diária máxima e, portanto, a maturidade sexual.

This study describes the morphological and functional analysis of spermatogenesis in guinea pigs (Cavia porcellus) with five (W5), six (W6), nine (W9) and eleven (W11) weeks of age (n=5/group). The aspects analyzed include counts of cell populations present in stage 1 of seminiferous epithelium cycle (SEC), efficiency of spermatogonial mitosis (EMi), meiotic production (EMe), overall yield of spermatogenesis (EOS), Sertoli cell index (SCI) and carrying capacity of Sertoli cells (CCSC). The results showed that the average number of spermatogonia type A, primary spermatocytes in pré-leptóteno/leptóteno, primary spermatocytes in pachytene, total spermatogenic cells and Sertoli cells showed numerical variations according to age; however they were statistically not detected, while round spermatids increased significantly at puberty and then stabilized. The spermatogenic production of 5 to 11-week-old guinea pigs did not reach the stabilization point, and the RMi, RME, EOS, SCI and CCSC showed significant number variation as a function of age. The results demonstrate that Cavia porcellus in post-pubertal stage 2 are an advantageous experimental model to address studies on the processes of homologous recognition, alignment, and synapsis during meiotic prophase; intrinsic yield of spermatogenesis in guinea pigs is similar to Wistar rats, paca and agouti (Dasyprocta sp.) and lower than in cavies, whereas the functional efficiency of Sertoli cells is higher than in agouti and Wistar rats, and lower than in pacas, spiny rat and collared peccaries. We conclude that in guinea pigs the spermatogenesis is fully established at 6 weeks of age, indicating the pubertal stage of sexual development, and until week 11 they do not reach the maximum daily sperm production and therefore sexual maturity.

Morphological, stereological and histometrical assessment of the testicular parameters between holstein and simmental bulls / Evaluación morfológica, estereológica e histométrica de los parámetros testiculares entre toros holstein y simmental

The testicular measurement is important criteria for experimental researches especially in toxicological studies and the prediction of spermatogenesis. In light of this knowledge, we aimed to estimate and compare these parameters in two different kinds of cattle breeds. The gross anatomical measurements were performed by vernier caliper, volume of the testis was estimated by Cavalieri method and seminiferous tubule diameter was measured on the histological sections by software-loaded computer attached to microscope. The mean testis weight, length, width, volume, and tubule diameter of the Simmental bulls and the Holstein bulls measured as 316 g, 12.1 cm, 6.9 cm, 295 cm3 and 226.68 um and 299 g, 12.1 cm, 6.8 cm, 285 cm3 and 223.44 µm, respectively. In conclusion all investigated parameters were not found statistically important in groups and between the breeds (p>0.05). The authors believed that the obtained data will contribute to the literature and facilitate future research.

La medición testicular es un criterio importante para las investigaciones experimentales sobre todo en los estudios toxicológicos y predicción de la espermatogénesis. El objetivo fue estimar y comparar estos parámetros en dos diferentes tipos de razas de ganado. Las mediciones anatómicas fueron realizadas con un pie de metro, el volumen de los testículos se estimó por el método de Cavalieri y el diámetro de los túbulos seminíferos se midió en las secciones histológicas observadas al microscopio mediante software. La Media del peso testicular, longitud, ancho, volumen y diámetro de los túbulos seminíferos de los toros Simmental fueron 316 g, 12,1 cm, 6,9 cm, 295 cm3 y 226,68 µm y de los toros Holstein fueron 299 g, 12,1 cm, 6,8 cm, 285 cm3 y 223,44 um, respectivamente. En conclusión, todos los parámetros investigados no tuvieron una importante significación en los grupos y entre las razas (p> 0,05). Creemos que los datos obtenidos contribuirán a la literatura y facilitar las futuras investigaciones.

Effects of cypermethrin on male reproductive system in adult rats / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To evaluate effects of cypermethrin on the testis histology and testosterone, LH and FSH in adult male Sprague-Dawley rats.</p><p><b>METHODS</b>The intact adult male rats were randomly divided into five groups and were treated with cypermethrin at doses of 0, 7.5, 15, 30, or 60 mg/kg per day by oral gavage for 15-days. After the treatments, serum was collected for hormone assays. The testes, epididymides, seminal vesicles, and prostates were excised and weighed. The right testis was frozen for daily sperm production and the left one was processed for histopathology.</p><p><b>RESULTS</b>Daily sperm production decreased significantly in 30 and 60 mg/(kg•day) groups. Testicular structure abnormalities included atrophic and distorted seminiferous tubules, deformed and disordered arrangement of germ cells, reduced germ cells, Sertoli cells and Leydig cells, vacuolization and multinucleated formations of spermatids in the cypermethrin-treated rats. Vacuolization was found in Sertoli cells and the deformed nucleus was noted in Leydig cells. Serum testosterone reduced significantly in 30 and 60 mg/(kg•day) groups. Serum FSH increased significantly in 60 mg/(kg•day) group.</p><p><b>CONCLUSION</b>Cypermethrin induces impairments of the seminiferous tubules structure and spermatogenesis in the rats. The damages of the male reproductive system may be attributed to the imbalance of circulating testosterone.</p>

Microscopia e morfometria de componentes tubulares de testículos de coelhos suplementados com geleia real / Microscopy and morphometry of tubular components of rabbit testis supplemented with royal jelly

Estudaram-se os efeitos da geleia real sobre a espermatogênese de coelhos tratados com diferentes concentrações de geleia real. Os tratamentos foram formados por três grupos: grupo-controle; grupo que recebeu 0,5mg/dia de geleia real; e grupo que recebeu 1,0mg/dia de geleia real. O estudo envolveu a morfometria testicular. Não houve diferença entre os tratamentos quanto aos pesos corporal (T1=3,20±0,19kg, T2=2,96±0,30kg e T3=3,21±0,37kg) e gonadal (T1=2,36±0,33g, T2=2,53±0,33g e T3=2,64±0,39g) e quanto aos índices gonadossomático (T1=0,15±0,02%, T2=0,17±0,03% e T3=0,16±0,02%) e tubulossomático (T1=0,06±0,01%; T2=0,07±0,01% e T3=0,06±0,01%). O diâmetro médio dos túbulos seminíferos (T1=225,95±13,27µm, T2=239,68±21,50µm e T3=231,57±15,94µm), a altura do epitélio seminífero (T1=66,05±5,37µm, T2= 73,47±9,11µm e T3=63,34±4,79µm) e o comprimento de túbulos seminíferos por testículo (T1=46,63±13,44m, T2=43,58±12,17m e T3=46,96±9,54m) e por grama de testículo (T1=19,50±2,68m, T2=17,12±3,91m e T3=17,78±1,98m) não diferiram entre tratamentos. Conclui-se que a suplementação com geleia real, nas doses utilizadas, não altera as características testiculares avaliadas.

This study aimed to investigate the effects of royal jelly on spermatogenesis in rabbits treated with different concentrations of RJ (Control; 0,5mg/day; and 1,0mg/day) using testicular morphometry. There was no significant difference between the body weight (T1= 3.20±0.19kg; T2= 2.96±0.30kg; T3=3.21±0.37kg) and gonadal weight (T1= 2.36±0.33g; T2= 2.53±0.33g; T3= 2.64±0.39g), gonadossomatic index (T1= 0.15±0.02%; T2= 0.17±0.03%. T3= 0.16±0.02%) and tubulossomatic index (T1= 0.06±0.01%; T2= 0.07±0.01%. T3= 0.06±0.01%) between treatments, showing that the percentage of body mass, and the percentage of seminiferous tubules allocated in testis were similar in the 3 experimental groups. Similarly, the mean diameter of the seminiferous tubules (T1= 225.95±13.27µm; T2=239.68±21.50µm; T3= 231.57±15,94µm), the height of the seminiferous epithelium (T1=66,05±5,37µm; T2=73.47±9.11µm; T3=63.34±4.79 µm) and length of seminiferous tubule for testis (T1=46.63±13.44m; T2=43.58±12.17m; T3=46.96±9.54m) and per gram of testis (T1=19.50±2.68m; T2=17.12±3.91m; T3=17.78±1.98m) did not differ statistically. It was concluded that supplementation with royal jelly, at the doses used, did not alter the testicular parameters evaluated here.

Testicular morphology and cauda epididymal sperm reserves of male rats exposed to Nigerian Qua Iboe Brent crude oil

Potential negative effects of exposure to Nigerian Qua Iboe Brent crude oil on the reproductive system of male rats was investigated. Forty Sprague-Dawley rats were used for the experiment. Exposure to Nigerian Qua Iboe Brent crude oil was achieved via oral administration of increasing doses (0.1, 0.2, and 0.4 ml/rat) every other day for 4 weeks. Cauda epididymal sperm reserves and relative weights of the testes as well as histological features of the testes of rats that received the crude oil treatment were compared to those of control rats. The results described here showed a significant (p < 0.01) dosedependent reduction in the cauda epididymal sperm reserves of rats that received crude oil treatment relative to the control group. The morphology of testes of the crude oil-exposed rats was characterized by the presence of interstitial exudates, degeneration, and necrosis of spermatogenic and interstitial (Leydig) cells. Findings indicate that exposure of male rats to Nigerian Qua Iboe Brent crude oil may have adversely affected their reproductive systems. This may imply possible reproductive health hazards for animals and humans that may be exposed to this environmental pollutant, especially in areas where oil spillage is a common feature.

Light and Electron Microscopic Observation in the Frozen-thawed Mouse Testicular Tissues / 대한불임학회지

OBJECTIVE: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. METHODS: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. RESULTS: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. CONCLUSIONS: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

Light and Electron Microscopic Observation in the Frozen-thawed Mouse Testicular Tissues / 대한불임학회지

OBJECTIVE: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. METHODS: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. RESULTS: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. CONCLUSIONS: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

Semen cuscutae flavonoids protect cells of rat seminiferous tubule from apoptosis induced by serum withdraw / 中国药理学通报

Aim To study the effect of Semen Cuscutae flavonoids on oxidative damage and apoptosis of rat seminiferous tubule cells. Methods Detecting apoptosis cells was performed by TUNEL methods and oxidative damage index such as MDA, T-AOC and ROS. Results Semen Cuscutae flavonoids could inhibit testis cells oxidative damage and apoptosis, the effect is dose dependent and the most significantly in 500 mg?L -1. Conclusion Semen Cuscutae flavonoids are effective antioxidants, which can inhibit apoptosis and oxidation of testis cells.

THE EFFECT OF ALCOHOL ON THE EXPRESSION OF eNOS,PCNA AND CELL APOPTOSIS OF SEMINIFEROUS TUBULES IN THE MOUSE TESTES / 解剖学报

Objective To investigate the change of eNOS,the spermatogenic cell proliferation,apoptosis in mouse testis exerted by alcohol. Methods The immunohistochemical staining method for detecting of eNOS,PCNA,TUNEL method for detecting of apoptotic cells and the satistics analysis were used in the present study. Results With the increase of alcohol concentration,the structure of seminiferous tubule changed,the diameter of seminiferous tubule decreased,the surface density of postive eNOS cells increased gradually,and the number of positve PCNA cells and apoptosis cells also increased.There were significant difference in 15% alcohol concentration group compared with the other groups(P

Testes biopsy analysis and hormonal reproduction levels of pig tail macaque (Macaca nemestrina) injected with testosterone enanthate (TE) and depot medroxy progesterone acetate (DMPA).

The objective of this study is to analyse the testes biopsy and to know the lnrmone level reproduction of pig tail macaque (Macaca nemestrina) injected with testosterone enanthate (TE) and depot medroxyprogesterone acetate (DMPA). Six pig tail male macaques, age 6 - 8 years old were used as sanrple. Three months after adaptation period, each animal was injected intra muscularly with 32 mg TE each week starting at week zero up to the sixth week. The treatment was continued every 3 weeks after the sixth week up to the 24th week. 40 mg of DMPA was injected intramuscularly at week zero, and continued every week up to week 18. Volume of the testes was taken every three weeks and blood samples for examination of gonadotropin hormone and steroid hormone were taken at 6 week intervals. Testes biopsy was perfomed at week 30 and week 48. Preparation of testes histological slides were made using the paraffin method and stained with hematoxylin-Eosin (HE). The results of this study showed that both testes volume decreased i.e. 18.35 cm3 ± 9.35 and 19.02cm3 ± 10.88 (at week zero) to 6.70 cm3 ± 3.80 and 7.02 cm3 ± 4.61 (lowest volume at week 21). In recovery period, the testes volume increased to 20.34 cmr ± 7.87 and 21.75 cm3 ± 7.09. The diameter of the seminiferous tubules and the score of spermatogenesis (right and left testes) at week 30 were 0.13 mm ± 0.027 and 0.13 mm ± 0.026 and score were 5.08 ± 2.67 and 5.41 ± 2.51. At week 48, both diameter of seminiferous tubules and spermatogenesis score increased to become 0.18 mm ± 0.029 and 0.18 mm ± 0.026, and score were 7.51 ± 2.14 and 7.57 ± 1.59, During this period, hyalinization and fibrosis of seminiferous tubule occurred. By week 6, the total testosterone, free testosterone, and estrogen hormone levels increased quite sharply and then decreased but still higher than base levels of hormone. In the recovery period, estrogen hormone increased significantly until the end of observation (week 48). FSH and LH hormone levels decreased until week 6, then the FSH hormone levels increased until the end of observation, while the LH hormone level is still lower than base level. Conclusion of this study is the injection of TE and DMPA combination will alter the histological structure of the pig tailed macaque testes i.e. decreasing the diameter of the seminiferous tubules, suppressing spermatogenesis and hyalinisation and fibrosis of seminiferous tubules. The damages of this structure are likely caused by inhibition of feedback mechanism of hypothalamus-hypophysis-testes.

Influence on the Pregnancy Rate of Motility of Fresh and Frozen Testicular Spermatozoa in Obstructive Azoospermic Patients / 대한불임학회지

OBJECTIVE: ICSI with testicular sperm could achieve optimal fertilization and pregnancy. This study was performed to observe the influence on fertilization and pregnancy of motility of fresh testicular sperm and sperm extracted from frozen-thawed seminiferous tubules in obstructive azoospermia. MATERIALS ANDMETHODS: We analysed clinical outcome of ICSI using fresh testicular sperm and sperm extracted from thawed seminiferous tubules. The presence of motility were compared to determine the factor for optimal fertilization and pregnancy rates. RESULTS: In 316 cases of TESE-ICSI in obstructive azoospermia, ICSI with fresh testicular sperm (fresh sperm group) were 163 cases and ICSI with sperm testicular sperm extracted from frozen-thawed seminiferous tubule (thawed sperm group) were 153 cases. The fertilization rates were 71.3% and pregnancy rates were 32.5% in fresh sperm group, in thawed sperm group, 65.1% and 33.3% respectively. The fertilization and pregnancy rates of motile and non-motile testicular sperm were 72.9% and 33.6%, 50.0% and 18.2%, respectively (p<0.05). The fertilization and pregnancy rates of motile and non-motile sperm extracted from the thawed seminiferous tubule were 67.8% and 34.7%, 55.1% and 28.1%, respectively (p<0.05). The comparative of the results of ICSI using motile fresh testicular sperm and motile sperm extracted from thawed seminiferous tubule, fertilization and pregnancy rates were not significantly different (72.9% and 33.6%, 67.8% and 34.7%, respectively). CONCLUSION: These results suggest that successful pregnancy in TESE-ICSI treatment is influenced by the motility of fresh testicular sperm and sperm extracted from thawed seminiferous tubule in obstructive azoospermic patients.

Histological and Biochemical Study on the Effect of Cadmium Chloride on the Rat Testis / 대한해부학회지

Although cadmium is a well known heavy metal which has an influence testis and brings about male infertility, the mechanism of action in the testis is still fully unknown. In these experiment, cadmium chloride 4 mg/kg of body weight administered intraperitoneally to the rat (Sprague-Dawley) and sacrificed after 1 week, and morphological changes were observed by LM and TEM. In addition, electrophoresis, immunoprecipitation and Western blotting and N-terminal analysis performed to reveal the protein changes. 1. Major findings under light microscope were hemorrhagic necrosis and death of all the spermatogenic cells and supporting cells within the seminiferous tubules, and decreased volume of ECM, many apoptotic bodies, and death of interstitial cells and fibroblasts within interstitium. 2. The EM findings were disruption of nuclear membrane and disappearance of cell organelles of spermatogenic cells and supporting cells within seminiferous tubules, and decreased filopodia, increased inclusion bodies, vacuolation and apoptotic changes of the interstitial cells and fibroblastic cells, many short electron-dense collagen fibers in the extracellular matrix of interstitium. 3. Two proteins of molecular weight 42 kDa and 21 kDa which disappeared after cadmium treatment were rat collagen type I alpha 2. According to the above results, it is considered that cadmium degrades the collagen of the wall of small blood vessels within seminiferous tubules and interstitium and disrupts vascular walls, which results hemorrhagic necrosis, death of all the spermatogenic cells, and the death of interstitial cells and fibroblastic cells.